This invention relates to a medicament, particularly a pyrazole derivative having an action to inhibit calcium release-activated calcium channel, and a pharmaceutical composition containing the same as an active ingredient, particularly a calcium release-activated calcium channel inhibitor.
It has been known for a long time that calcium ion (Ca2+) is important for an intracellular second messenger in the activation of various cells. Intracellular Ca2+ also acts as an important regulatory factor in inflammatory cells. It has been suggested, however, that voltage-operated Ca2+ channel (to be referred to as xe2x80x9cVOCCxe2x80x9d hereinafter) inhibitors such as nifedipine does not show inhibitory activity against the activation of inflammatory cells and that a Ca2+ influx mechanism other than VOCC exist in inflammatory cells.
Hoth et al. have cenorted that Ca2+-selecive and Ca2+ store depletion-activated Ca2+ channel, namely Ca2+ released-activated Ca2+ channel (to be referred to as xe2x80x9cCRACCxe2x80x9d hereinafter; also called store-dependent Ca2+ channel), is present in mast cells and lymphocytes, and these cells are insensitive to membrane potential (Pflugers Arch., 430, pp. 315-322 (1995)). It is known that CRACC is present in several inflammatory cells such as mast cells, lymphocytes, astrocytes (J. Biol. Chem., 270, pp. 29-32 (1995)) and the like, and that it is deeply concerned in, for example, cytokine production and lipid mediator release (J. Immunol., 155, pp. 285-296 (1995) and Br. J. Pharmacol., 114, pp. 598-601 (1995)).
Recently, it has been revealed that tenidap, an agent for treating rheumatoid arthritis, has a potency of CRACC inhibitor (Cell Calcium, 14, pp. 1-16 (1993)). Therefore, a CRACC inhibitor has a possibility of therapeutic potency on chronic inflammatory diseases including rheumatoid arthritis.
It is known that CRACC is also present in endothelial cells (Am. J. Physiol., 269, C 733-738 (1995)) and epithelial cells (J. Biol. Chem., 270, pp.29169-175 (1995)). Since it has been reported that sustained calcium influx takes a role in the radical affection of endothelial cells (Am. J. Physiol., 261, C 889-896 (1991)), it is suggested that a CRACC inhibitor should have protective efficacy on endothelial cell-concerned tissue damage.
In addition, it has been reported that blockades of calcium influx inhibit cell proliferation and interleukin 2 (IL-2) production (Br. J. Pharmacol., 113 , pp. 861-868 (1994)). Therefore, a CRACC inhibitor is useful as an agent for the prevention and treatment of proliferative or progressive diseases (e.g., malignant tumor and the like) and autoimmune diseases, and also as a suppresser for tissue rejection in transplantation.
On the other hand, it is known that in excitable cells such as smooth muscle cells and nerve cells, intracellular calcium is mainly regulated with VOCC not with CRACC. Therefore, it is expected that a calcium channel blocker having CRACC selectivity against VOCC should be an useful agent for the prevention or treatment of various inflammatory diseases, allergic diseases, autoimmune diseases, tissue damages, proliferative diseases and the like without undesirable actions on cardiovascular and central nervous system.
Recently, some compounds showing CRACC inhibitory activity have been reported, such as a cycloalkyl-piperazinylethanol derivative disclosed in a published German patent application 4404249 and a 2-(3,4-dihydro-1-isoquinolyl)acetamide derivative disclosed in WO 94/00435. It has also reported that 5-amino-1-[[3,5-dichloro-4-(4-chlorobenzoyl)phenyl]methyl]-1H-1,2,3-triazole-4-carboxamide inhibits CRACC (J. Pharm. Exp. Ther., 257, pp. 967-971 (1991)). However, there are no reports on a compound whose CRACC selectivity over VOCC has been confirmed.
On the other hand, a published German patent application 2525024 discloses a 5-(heterocycloylaminophenyl)-1-phenylpyrazole derivative which shows an anti-inflammatory activity. However, this patent does not disclose or suggest about its inhibitory activities against CRACC and IL-2 production.
WO 95/18097 discloses an anthranilic acid derivative represented by the following formula, which inhibits a cyclic GMP phosphodiesterase. In the formula, R1 to R4 represent H, a halogen atom, . . . , pyrazolyl which may be substituted, . . . ; n is 0 to 6, W represents N or CH, Y represents O or S, . . . (see said published patent application for details). 
An unexamined published Japanese patent application 9-59236 discloses an R1, R2-di-substituted benzamide derivative represented by the following formula, which is useful for the prevention and treatment of rheumatic, allergic and other inflammatory diseases. In the formula, R1 represents a substituted or unsubstituted aromatic heterocyclic ring, . . . , R2 represents a halogen, a nitro, xe2x80x94NR5R6, . . . , A represents xe2x80x94C(xe2x95x90Z) NR3R4 or xe2x80x94NR4C(xe2x95x90Z)R3, R3 represents a substituted or unsubstituted aromatic hydrocarbon ring, a substituted or unsubstituted aromatic heterocyclic ring . . . (see said published patent application for details). However, there is no illustrative disclosure about pyrazolyl as the aromatic heterocyclic ring group. In addition, there is no disclosure about inhibitory activities against CRACC and/or IL-2 production. 
The inventors of the present invention have conducted extensive studies on the screening of compounds having excellent CRACC inhibitory activity. As a result of the efforts, certain pyrazole derivatives which possess entirely different structures from those of the reported CRACC inhibitors have been found to show excellent CRACC inhibitory activity. The present invention has been accomplished by further finding that these compounds have high CRACC selectivity over VOCC.
Accordingly, the invention relates to a novel pyrazole derivative represented by the following general formula (I) which is characterized in that it has a pyrazolyl group unsubstituted or substituted with a specified group, or a pharmaceutically acceptable salt thereof. In the specification of this application, lower alkyl and halogen atom are abbreviated as Alk and Hal, respectively. 
(In the formula, each symbol has the following meaning:
D: pyrazolyl which may have 1 to 3 substituents selected rom the group consisting of -Alk, -lower alkenyl, -lower alkynyl, -halogeno-lower alkyl, -Alk-cycloalkyl, -Alk-O-Alk, -cycloalkyl, xe2x80x94O-Alk, xe2x80x94COOH, xe2x80x94COO-Alk and -Hal,
n: 0 or 1,
B: phenylene, a nitrogen-containing, divalent, saturated ring group, or a monocyclic, divalent heteroaromatic ring group which may be substituted with Alk,
X: xe2x80x94NR1xe2x80x94CR2R3xe2x80x94, xe2x80x94CR2R3xe2x80x94NR1xe2x80x94, xe2x80x94NR1xe2x80x94SO2xe2x80x94, xe2x80x94SO2xe2x80x94NR1xe2x80x94 or xe2x80x94CR4xe2x95x90CR5xe2x80x94,
R1: xe2x80x94H, xe2x80x94OH, -Alk, xe2x80x94O-Alk or xe2x80x94CO-Alk,
R2 and R3: the same or different from each other and each represents xe2x80x94H or -Alk, or R2 and R3 together form xe2x95x90O or xe2x95x90S,
R4 and R5: the same or different from each other and each represents xe2x80x94H, -Hal, -halogeno-lower alkyl or -Alk, and
A: benzene ring which may have one or more substituents; mono-, di- or tricyclic fused heteroaryl which may have one or more substituents; cycloalkyl which may have one or more substituents; a nitrogen-containing, saturated ring group which may have one or more substituents; lower alkenyl which may have one or more substituents; lower alkynyl which may have one or more substituents; or Alk which may have one or more substituents, or A and X may together form a group represented by a formula 
(wherein A2 is a nitrogen-obtaining hetero ring selected from the group consisting of 1-pyrrolidinyl, pyrazolidinyl, piperidino, piperazinyl, morpholino, 3,4-dihydro-2H-1,4-benzoxazin-4-yl and indolinyl, wherein the hetero ring may have one or more subsuituents), with the proviso that
(1) when D is 3,5-bis(trifluoromethyl)-1H-pyrazol-1-yl, n is 0, B is 1,4-phenylene and X is NHCO, A is a group other than 4-methyl-1,2,3-thiadiazol-5-yl,
(2) when D is 1-methyl-5-trifluoromethyl-1H-pyrazol-3-yl, n is 0, B is thiophene-2,5-diyl and X is CONH, A is a group other than 4-chlorophenyl,
(3) when D is 1-methyl-3-trifluoromethyl-1H-pyrazol-5-yl, n is 0, B is thiophene-2,5-diyl and X is CONH, A is a group other than benzyl,
(4) when D is 4-ethoxycarbonyl-5-trifluoromethyl-1H-pyrazol-1-yl, n is 0, B is 1,4-phenylene and Y is NHCO, A is a group other than trichlorovinyl,
(5) when D is 1H-pyrazol-1-yl, n is 0, B is 1,4-phenylene and Y is NHCO, A is a group other than 2-ethoxyvinyl, and
(6) when n is 1, D is 1H-pyrazol-5-yl substituted with at least one triflluoromethyl group or 1H-pyrazol-1-yl substituted with at least one trifuoromethyl group. The same shall apply hereinafter.)
The invention also relates to a pharmaceutical composition, particularly a pharmaceutical composition for use in the inhibition of calcium release activated calcium channel, which comprises a pyrazole derivative represented by the following general formula (Ixe2x80x2) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. Preferably, it relates to an IL-2 production inhibitor, a preventive or therapeutic agent for allergic, inflammatory or autoimmune diseases and a preventive or therapeutic agent for bronchial asthma or rheumatoid arthritis. 
(In the formula, each symbol has the following meaning:
D: pyrazolyl which may have 1 to 3 substituents selected from the group consisting of -Alk, -lower alkenyl, -lower alkynyl, -halogeno-lower alkyl, -Alk-cycloalkyl, -Alk-O-Alk, -cycloalkyl, xe2x80x94O-Alk, xe2x80x94COOH, xe2x80x94COO-Alk and -Hal,
n: 0 or 1,
B: phenylene, a nitrogen-containing, divalent, saturated ring group, or a monocyclic, divalent heteroaromatic ring group which may be substituted with Alk,
X: xe2x80x94NR1xe2x80x94CR2R3xe2x80x94, xe2x80x94CR2R3xe2x80x94NR1xe2x80x94, xe2x80x94NR1xe2x80x94SO2xe2x80x94, xe2x80x94SO2xe2x80x94NR1xe2x80x94 or xe2x80x94CR4xe2x95x90CR5xe2x80x94,
R1: xe2x80x94H, xe2x80x94OH, -Alk, xe2x80x94O-Alk or xe2x80x94CO-Alk,
R2 and R3: the same or different from each other and each represents xe2x80x94H or -Alk, or R2 and R3 together form xe2x95x90O or xe2x95x90S,
R4 and R5: the same or different from each other and each represents xe2x80x94H, -Hal, -halogeno-lower alkyl or -Alk, and
A: benzene ring which may have one or more substituents; mono-, di- or tricyclic fused heteroaryl which may have one or more substituents; cycloalkyl which may have one or more substituents; a nitrogen-containing, saturated ring group which may have one or more substituents; lower alkenyl which may have one or more substituents; lower alkynyl which may have one or more substituents or Alk which may have one or more substituents, or A and X may together form a group represented by a formula 
(wherein A2 is a nitrogen-containing hetero ring selected from the group consisting of 1-pyrrolidinyl, pyrazolidinyl, piperidino, piperazinyl, morpholino, 3,4-dihydro-2H-1,4-benzoxazin-4-yl and indolinyl, wherein the hetero ring may have one or more substituents), with the proviso that when n is 1, D is 1H-pyrazol-5-yl substituted with at least one trifluoromethyl group or 1H-pyrazol-1-yl substituted with at least one trifluoromethyl group. The same shall apply hereinafter.)
The following known compounds are included in the aforementioned general formula (Ixe2x80x2).
(1) A compound wherein D: 3,5-bis(trifluoromerhyl)-1H-pyrazol-1-yl, n: 0, B: 1,4-phenylene, Y: NHCO and A: 4-methyl-1,2,3-thiadiazol-5-yl (to be referred to as compound A hereinafter),
(2) a compound wherein D: 1-methyl-5-trifluoromethyl-1H-pyrazol-3-yl, n: 0, H.: thiophene-2,5-diyl, Y: CONH and A: 4-chlorophenyl (to be referred to as compound B hereinafter),
(3) a compound wherein D: 1-methyl-3-trifluoromethyl-1H-pyrazol-5-yl, n: 0, B: thiophene-2,5-diyl, Y: CONH and A: benzyl (to be referred to as compound C hereinafter),
(4) a compound wherein D: 4-ethoxycarbonyl-5-trifluoromethyl-1H-pyrazol-1-yl, n: 0, B: 1,4-phenylene, Y: NHCO and A: trichlorovinyl (to be referred to as compound D hereinafter), and
(5) a compound wherein D: 1H-pyrazol-1-yl, n: 0, B: 1,4-phenylene, Y: NHCO and A: 2-ethoxyvinyl (to be referred to as compound E hereinafter).
However, though the compounds A to D are described in the MAYBRIDGE""s reagent catalog (UK, Cornwall, published in August, 1995) as SEW04225, KM02940, KM03000 and GK02421, there are no reports on their application to not only medicaments as a matter of course but also other use. Also, the compound E is disclosed as a production material of a medicament in JP-A-61-82, but there is no description regarding its pharmacological actions. Thus, the pharmaceutical compositions which contain these known compounds are novel.
Preferred compounds of the general formula (I) or (Ixe2x80x2) of the Invention are pyrazole derivatives or pharmaceutically acceptable salts thereof, in which the pyrazoyl group of D is pyrazolyl (particularly 1H-pyrazol-5-yl or 1H-pyrazol-1-yl) which is substituted with at least trifluoromethyl group.
Other preferred compounds of the invention are listed below.
Pyrazole derivatives or pharmaceutically acceptable salts thereof, in which
1) A is phenyl which may have one or more substituents of F group; mono-, di- or tricyclic fused heteroaryl which may have one or more substituents of F group; cycloalkyl which may have one or more substituents of F group; a nitrogen-containing, saturated ring group which may have one or more substituents of F group; lower alkenyl which may have one or more substituents of G group; lower alkynyl which may have one or more substituents of G group; or Alk which may have one or more substituents of G group, wherein
the F group is a group consisting of -Alk, -lower alkenyl, -lower alkynyl, -Hal, xe2x80x94NH2, xe2x80x94NH(Alk), xe2x80x94N(Alk)2, xe2x80x94NO2, xe2x80x94CN, xe2x80x94OH, xe2x80x94O-Alk, xe2x80x94Oxe2x80x94CO-Alk, xe2x80x94SH, xe2x80x94S-Alk, xe2x80x94COOH, xe2x80x94COO-Alk, xe2x80x94CO-Alk, xe2x80x94CHO, xe2x80x94CONH2, xe2x80x94CONH(Alk), xe2x80x94CON(Alk)2, xe2x80x94SO-Alk, xe2x80x94SO2-Alk, xe2x80x94SO2NH2, xe2x80x94SO2NH-(Alk), xe2x80x94SO2N(Alk)2, -aryl, -cycloalkyl, xe2x80x94O-Alk-Oxe2x80x94, -halogeno-lower alkyl, -Alk-NH2, -Alk-NH(Alk), -Alk-N(Alk)2, -Alk-OH, -Alk-O-Alk, -Alk-SH, -Alk-S-Alk, -Alk-COOH, -Alk-COO-Alk, -Alk-CO-Alk, -Alk-CHO, -Alk-CONH2, -Alk-CONH(Alk), -Alk-CON(Alk)2, -Alk-SO-Alk, -Alk-SO2-Alk, -Alk-SO2NH2, -Alk-SO2NH(Alk), -Alk-SO2N(Alk)2, -Alk-aryl and -Alk-cycoalkyl, and the G group is a group consisting of -Hal, xe2x80x94NH2, xe2x80x94NH(Alk), xe2x80x94N(Alk)2, xe2x80x94NO2, xe2x80x94CN, xe2x80x94OH, xe2x80x94O-Alk, xe2x80x94Oxe2x80x94CO-Alk, xe2x80x94SH, xe2x80x94S-Alk, xe2x80x94COOH, xe2x80x94COO-Alk, xe2x80x94CO-Alk, xe2x80x94CHO, xe2x80x94CONH2, xe2x80x94CONH(Alk), xe2x80x94CON(Alk)2, xe2x80x94SO-Alk, xe2x80x94SO2-Alk, xe2x80x94SO2NH2, xe2x80x94SO2NH-(Alk), xe2x80x94SO2N(Alk)2, aryl which may have one or more substituents of F group; mono-, di- or tricyclic fused heteroaryl which may have one or more substituents of F group; cycloalkyl which may have one or more substituents of F group and a nitrogen-containing, saturated ring group which may have one or more substituents of F group, or A and X may together form a group represented by a formula 
(wherein A2 is a nitrogen-containing hetero ring selected from the group consisting of 1-pyrrolidinyl, pyrazolidinyl, piperidino, piperazinyl, morpholino, 3,4-dihydro-2H-1,4-benzoxazin-4-yl and indolinyl, wherein the hetero ring may have one or more substituents of F group),
2) B is phenylene; piperidine-1,4-diyl; or a monocyclic, divalent heteroaromatic ring selected from the group consisting of thiophene, furan, pyrrole, imidazole, pyrazole, thiazole, isothiazole, oxazole, isoxazole, thiadiazole, pyridine, pyrazine, pyridazine and pyrimidine, which may be substituted with Alk, X is xe2x80x94NHxe2x80x94COxe2x80x94, xe2x80x94NHxe2x80x94CH2xe2x80x94, xe2x80x94N(OH)xe2x80x94COxe2x80x94, xe2x80x94N(Alk)xe2x80x94COxe2x80x94, xe2x80x94COxe2x80x94NHxe2x80x94, xe2x80x94CH2xe2x80x94NHxe2x80x94, xe2x80x94COxe2x80x94N(OH)xe2x80x94, xe2x80x94COxe2x80x94N(Alk)-, xe2x80x94SO2NHxe2x80x94, xe2x80x94NHSO2xe2x80x94 or xe2x80x94CHxe2x95x90C(Hal)-, A is aryl which may have one or more substituents of group F; mono-, di- or tricyclic fused heteroaryl selected from the group consisting of thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, tetrazolyl, triazolyl, thiadiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, isoindolyl, isoquinolyl, quinolyl, quinoxanyl, phthalazinyl, imidazopyridyl, quinazolinyl and cinnolinyl, which may have one or more substituents of group F; cycloalkyl; a nitrogen-containing, saturated ring selected from the group consisting of pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidyl, piperazinyl and morpholinyl, which may be substituted with one or more Alk; lower alkynyl which may be substituted with one or more Hal; lower alkenyl which may be substituted with one or more Hal; or Alk which may be substituted with one or more Hal, and the F group is a group consisting of -Alk, -lower alkenyl, -lower alkynyl, -Hal, xe2x80x94NH2, xe2x80x94NH(Alk), xe2x80x94N(Alk)2, xe2x80x94NO2, xe2x80x94CN, xe2x80x94OH, xe2x80x94O-Alk, xe2x80x94Oxe2x80x94CO-Alk, xe2x80x94SH, xe2x80x94S-Alk xe2x80x94COCH, xe2x80x94COO-Alk, xe2x80x94CO-Alk, xe2x80x94CHO, xe2x80x94CONH2, xe2x80x94CONH(Alk), xe2x80x94CON(Alk)2, xe2x80x94SO-Alk, xe2x80x94SO2-Alk, xe2x80x94SO2NH2, xe2x80x94SO2NH-(Alk) and xe2x80x94SO2N(Alk)2, or A and X may together form a group represented by a formula 
3) n is 0, D is pyrazolyl which may have 1 to 3 substituents selected from -Alk, -halogeno-lower alkyl, xe2x80x94COOH and xe2x80x94COO-Alk, B is phenylene or a monocyclic, divalent heteroaromatic ring selected from the group consisting of thiophene, furan, thiazole, pyridine and pyrimidine, which may be substituted with Alk, X is xe2x80x94NHxe2x80x94COxe2x80x94, xe2x80x94N(OH)xe2x80x94COxe2x80x94, xe2x80x94COxe2x80x94NHxe2x80x94, xe2x80x94CH2xe2x80x94NHxe2x80x94 or xe2x80x94COxe2x80x94N(Alk)-, and A is phenyl which may have one or more substituents selected from the group consisting of -Alk, -Hal, xe2x80x94NH2, xe2x80x94N(Alk)2, xe2x80x94NO2, xe2x80x94CN, xe2x80x94OH, xe2x80x94O-Alk and xe2x80x94COO-Alk; mono-, di- or tricyclic fused heteroaryl selected from the group consisting of thienyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, tetrazolyl, triazolyl, thiadiazolyl, pyridyl, pyrazinyl and isoquinolyl, which may be substituted with Alk; cycloalkyl; lower alkenyl which may be substituted with one or more Hal; or Alk, or
4) X is xe2x80x94NHxe2x80x94COxe2x80x94 or xe2x80x94COxe2x80x94NHxe2x80x94.
Particularly preferred is a pyrazole derivative or a pharmaceutically acceptable salt thereof, in which D is 1-methyl-3-trifluoromethyl-1H-pyrazol-5yl and A is phenyl which may be substituted with Hal, or D is 3,5-bis(trifluoromethyl)-1H-pyrazol-1-yl and A is monocyclic heteroaryl selected From the group consisting of thiazolyl, thiadiazolyl, thienyl and pyridyl, which may be substituted with Alk.
Unless otherwise noted, the term xe2x80x9clowerxe2x80x9d as used herein means a straight or branched carbon chain having from 1 to 6 carbon atoms. Examples of preferred groups include methyl, ethyl, propyl and the like as xe2x80x9clower alkyl (Alk)xe2x80x9d, vinyl, 1-propenyl, 1,2-dimethyl-1-propenyl and the like as xe2x80x9clower alkenylxe2x80x9d and ethynyl and the like as xe2x80x9clower alkynylxe2x80x9d. The xe2x80x9chalogen atom (Hal)xe2x80x9d is I, Br, F or Cl. The xe2x80x9chalogeno-lower alkylxe2x80x9d is an Alk substituted with one or more Hal, and trifluoromethyl is particularly preferable. The xe2x80x9carylxe2x80x9d is an aryl group having from 6 to 14 carbon atoms, preferably phenyl or naphthyl. The xe2x80x9ccycloalkylxe2x80x9d is a cycloalkyl having from 3 to 8 carbon atoms, preferably cyclopropyl or cyclohexyl.
The xe2x80x9cmonocyclic, divalent heteroaromatic ring groupxe2x80x9d is a five- or six-membered monocyclic, divalent heteroaromatic ring group which contains from 1 to 4 hetero-atoms selected from N, S and O atoms, and furan-2,5-diyl, thiophene-2,5-diyl, thiazole-2,5-diyl, pyridine-2,3-diyl and pyrimidine-2,5-diyl are particularly preferable. The xe2x80x9cphenylenexe2x80x9d is preferably 1,4-phenylene.
The xe2x80x9cmono-, di- or tricyclic fused heteroarylxe2x80x9d is a five- or six-membered mono-, di- or tricyclic fused ring which contains from 1 to 5 of O, S or N atom as heterocyclic atoms. The xe2x80x9cnitrogen-containing saturated ringxe2x80x9d is a five- or six-membered nitrogen-containing saturated ring which contains 1 or 2 N atoms as ring atoms and may further contain one O or S atom. The xe2x80x9cnitrogen-containing, divalent, saturated ring groupxe2x80x9d is preferably piperidine-1,4-diyl. When n is 0, D and B are directly bonded.
The compound of this invention may exist in the form or geometrical isomers or tautomers depending on the kinds of substituent groups, and these isomers in separated forms or mixtures thereof are included in the present invention. Also, the compound of the present invention may have asymmetric carbon atoms, so that it may exist in (R) and (S) optical isomer forms based on such carbon atoms. All of the mixtures and the isolated forms of these optical isomers are included in the present invention.
The compound (I) or (Ixe2x80x2) of this invention may form an acid addition salt or, depending on the kinds or substituent groups, a salt with a base. Such salts are pharmaceutically acceptable ones, and their preferred examples include acid addition salts with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid and the like) or with organic acids (e.g., formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, aspartic acid, glutamic acid and the like) and salts with inorganic bases (e.g., sodium, potassium, magnesium, calcium, aluminum and the like) or with organic bases (e.g., methylamine, ethylamine, ethanolamine, lysine, ornithine and the like), as well as ammonium salts.
In addition, various hydrates and solvates and polymorphism of the compound (I) or (Ixe2x80x2) and salts thereof are also included in this invention.
(Production Method)
The compound of the present invention and a pharmaceutically acceptable salt thereof can be produced by making use of the features of its basic structure or the kinds of its substituents and by employing various known synthesis methods. In that case, depending on the kind of each functional group, it may sometimes be effective from the viewpoint of production techniques to replace said functional group with an appropriate protecting group, namely a group which can be converted into said functional group easily, at the stage of raw materials or intermediates. Thereafter, the compound of interest can be obtained by removing the protecting group as occasion demands. Examples of such functional groups include a hydroxyl group, a carboxyl group and the like and examples of their protecting groups include those which are described in xe2x80x9cProtective Groups in Organic Synthesisxe2x80x9d, 2nd edition, edited by Greene and Wuts, which may be optionally used depending on the reaction conditions.
The following describes typical methods for the preparation of the compound of the present invention. 
In this method, as shown in the above reaction formula, the compound (I-1) or (I-2) of the present invention is obtained by subjecting an amine derivative represented by the general formula (II) or (V) and a carboxylic acid derivative represented by the general formula (III) or (IV) to amidation reaction.
The carboxylic acid derivative (III) or (IV) which can be used in the He production method 1 is a free carboxylic acid or a reactive derivative thereof, and examples of the reactive derivative include acid halides such as acid chlorides, acid bromides and the like; acid azides; active esters which can be prepared using methanol, ethanol, benzyl alcohol, phenol which may be substituted, 1-hydroxybenzotriazole, N-hydroxysuccinimide and the like; symmetric acid anhydrides; and mixed acid anhydrides with alkylcarboxylic acid, p-toluencsulfonic acid and the like. These reactive derivatives are commercially available or can be produced by the usual procedures.
The amidating reaction can be carried out by the usual procedures.
When the reaction is carried out using a free carboxylic acid, it is necessary to use a condensing agent such as N,Nxe2x80x2-dicyclohexylcarbodiimide (DCC), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (WSCD) or the like or carboxylic acid activating agent such as 1,1xe2x80x2-carbonyldiimidazole, N,Nxe2x80x2-disuccinimhdyl carbonate, diphenylphosphoryl, azide, phosphorus oxychloride, phosphorus trichloride, triphenylphosphine/N-bromosuccinimide or the like.
The reaction is carried out using an amine derivative represented by the general formula (II) or (IV) and a carboxylic acid derivative represented by the general formula (III) or (IV), in equimolar amounts or one of them in excess amount, in a reaction inert organic solvent such as pyridine, tetrahydrofuran (THF), dioxane, ether, benzene, toluene, dichloromethane, 1,2-dichloroethane (DCE), chloroform, N,N-dimethylformamide (DMF), ethyl acetate, acetonitrile or the like. The reaction temperature is optionally selected depending on the kinds of reaction derivatives.
Depending on the kinds of reaction derivatives, addition of a base such as triethylamine, pyridine, picoline, N,N-dimethylaniline, potassium carbonate, sodium hydroxide or the like may be advantageous in some cases from the viewpoint of accelerating the reaction. It is possible to use pyridine also as the solvent. 
(In the above reaction formula, each of Ra and Rb represents H or Alk.)
In this production method, the compound (I-3) of the present invention is obtained by carrying out trifluoroacetylation of the carbon atom adjacent to the ketone of a compound represented by the general formula (VI) and then effecting cyclization by reacting it with a hydrazine derivative.
The first step trifluoroacetylation can be carried out by allowing the compound to react with a trifluoro acetylation agent (for example, ethyl trifluoroacetate, trifluoroacetic anhydride or the like) at a temperature of from xe2x88x9278xc2x0 C. to reflux temperature in a solvent such as methanol, ethanol, 1,3-dimethylimidazolidin-2-one (DMI), THF, DMF or the like, in the presence of a base such as sodium methoxide, sodium ethoxide, alkali metal hexamethyldisilazide, alkali metal hydride alkyl lithium, triethylamine or the like.
The second step cyclization reaction can be carried out by allowing the compound obtained in the first step to react with a hydrazine derivative in a solvent such as methanol, ethanol or the like, or without solvent, in the presence or absence of an acid such as acetic acid, hydrochloric acid or the like or Lewis acid such as titanium(IV) isocropoxide, titanium(IV) chloride, boron trifluoride-diethyl ether complex or the like. This reaction can be carried out at a temperature of from cooling temperature to reflux temperature. 
As shown in the above reaction formula, this production method is a method in which the compound (I-4) or (I-5) of the invention is obtained by a reductive amination reaction of an amine derivative represented by the general formula (II) or (V) with an aldehyde derivative represented by the general formula (VII) or (VIII).
This reductive amination reaction is carried out by allowing both compounds to react with each other in the same inert solvent of the case of amidation of the production method 1, and reducing the thus formed Schiff base after its isolation or directly without isolation. It is advantageous to carry out formation of Schiff base in the presence or the aforementioned Lewis acid, p-toluenesulfonic acid, adipic acid, acetic acid, hydrochloric acid or the like acid catalyst, in the presence of molcular sieves, potassium hydroxide or the like dehydrating agent or by removing formed water using Dean-Stark trap. The reaction temperature can be optionally set but is preferably from room temperature to reflux temperature.
Reduction of the Schiff base can be carried out at a temperature of from xe2x88x9220xc2x0 C. to heat reflux, by adding a reducing agent such as a metal hydride complex (e.g., sodium cyanoborohydride, sodium triacetoxyborohydride or sodium borohydride) or borane. Alternatively, it can be effected by carrying out the reaction using a reduction catalyst (e.g., palladium-carbon or Raney nickel) at a temperature of from 0xc2x0 C. to 100xc2x0 C. in a hydrogen atmosphere of from ordinary pressure to 50 kg/cm2, in a solvent such as methanol, ethanol, ethyl acetate, acetic acid or the like in the presence or absence of an acid such as acetic acid, hydrochloric acid or the like.
A compound of the invention in which X is xe2x80x94SO2xe2x80x94NR1xe2x80x94 or xe2x80x94NR1xe2x80x94SO2xe2x80x94 can be produced in the same manner as the aforementioned production method 1, except that a sulfonic acid derivative is used instead of the carboxylic acid derivative.
A compound in which X is xe2x80x94CR4xe2x95x90CR5xe2x80x94 can be produced by effecting formation of an olefin from an organic phosphorus compound and an aldehyde by the Horner-Emmons reaction or Witting reaction. This reaction can be carried out at a temperature of from xe2x88x9278xc2x0 C. to heat reflux in THF, DMF or the like solvent in the presence of a base such as lithium diisopropylamide, sodium hydride, triethylamine, alkyl lithium or phenyl lithium.
N-Alkylation of the nitrogen atom of amino group or amido group of X and N-alkylation of the ring nitrogen atoms can be carried out by a usually used N-alkylation method, for example, by allowing an amine derivative to react with an alkyl compound having a usual leaving group such as a halogen atom or an organic sulfone residue, at a temperature of from cooling to reflux in DMF, acetone, 2-butanone, acetonitrile or the like inert solvent or without solvent in the presence or absence of potassium carbonate, triethylamine, sodium hydride or the like base.
In addition to be above, introduction of substituents into respective rings, modification of groups, elimination of protecting groups and the like techniques can be carried out in the usual way.
Starting compounds of the aforementioned production methods are commercially available or can be produced easily by methods well known to those skilled in the art.
Each of the reaction products obtained by the aforementioned production methods is isolated and purified as a tree compound, a salt thereof, a hydrate thereof or a solvate thereon. The salt can be produced by a usual salt forming method. The isolation and purification are carried out by employing usually used chemical techniques such as extraction, concentration, evaporation, crystallization, filtration, recrystallization, various types of chromatography and the like. Various forms of isomers can be isolated by the usual procedures making use of physicochemical differences among isomers. For example, optical isomers can be separated by means of a conventional racemic resolution method such as fractional crystallization or a chromatography. In addition, an optical isomer can also be synthesized from an appropriate optically active starting compound.
The compound of the present invention is useful as an active ingredient of pharmaceutical compositions. Since it has inhibitory activities on CRACC and IL-2 Production, it is parcicularly useful as an inhibitor of CRACC or IL-2 production.
It also is particularly useful as an agent for use in the prevention and treatment of allergic, inflammatory or autoimmune diseases in which CRACC and/or IL-2 production are concerned. In this connection, examples of the allercic, inflammatory or autoimmune diseases include various diseases in which CRACC and/or IL-2 production are concerned such as bronchial asthma, psoriasis, atopic diseases including atopic dermatitis, inflammatory bowel diseases including Crohn disease, peptic ulcer, glomerular nephritis, hepatitis, pancreatitis, collagen disease, rheumatoid arthritis, osteoarthritis, rejection on transplantation and the like.
Applicability of the compound of the present invention to the aforementioned diseases is evident from the results of in vitro tests on inhibition of CRACC and IL-2 production, which will be described later, as well as the results of various tests carried out using animal models for diseases such as an antigen-induced airway eosinoohilia as a typical model for bronchial asthma, some T-cell-dependent disease models and a collagen-induced arthritis in mice. In addition, since the compounds of the present invention also have inhibitory effects on IL-4, IL-5, MMP-1 and TNFxcex1 production, such results also support its applicability to the aforementioned diseases.
On the other hand, anti-proliferative effect of the CPJCC inhibitor suggests that it should be useful in preventing or treating proliferative or progressive diseases such as malignant tumor, arteriosclerosis, multiple organ sclerosis, various types of fibrosis, burn keloid and the like. Also, since the CRACC inhibitor inhibits activation of inflammatory cells such as mast cells, leukocyoes and atrocities, which concern with inflammation in several peripheral or brain tissues, its action to protect tissues from their damages such as ischemia-reperfusion injury, head injury, cerebral infarction and myocardial infarction can be expected.
In particular, the compound of the present invention which is possessed of CRACC selective inhibitory activity over VOCC is useful, because it can cause CRACC inhibition without VOCC inhibition-induced undesirable reactions in central nervous system and cardiovascular system and the like.
The following shows certain tests and their results in order to confirm pharmacological actions of the compound of the present invention.
(1) CRACC Inhibitory Activity
Jurkat cells (6xc3x97106/ml) suspension loaded with a calcium indicator fluorescence dye fura-2 (1 xcexcM) was dispensed in 100 xcexcl portions into wells of a 96 well microplate. Intracellular calcium increase stimulated with a calcium pump inhibitor (thapsigargin) was induced by adding to each well a 100 xcexcl of Hanks"" balanced salt solution containing a drug to be tested in two times higher concentration than the final concentration and 2 xcexcM of thapsigargin (final concentration, 1 xcexcM), and, after 30 minutes of the addition, a fluorescence intensity ratio (R) was calculated from two fluorescence intensities obtained at excitation wave lengths of 340 nm/500 nm and 380 nm/500 nm, respectively. In calculating R, self-fluorescence of the drug to be tested was measured in a cell-free system, and the effect of the self-fluorescence on the fura-2 fluorescence was corrected.
The intracellular calcium concentration was obtained by the following calculation formula based on a maximum reaction of R (Rmax) obtained by 25 xcexcM ionomycin stimulation, a minimum reaction of R (Rmin) obtained by 5 xcexcM ionomycin+1 mM EGTA stimulation, a fluorescence efficiency (Sb2) of a calcium binding dye at an excitation wave length of 380 nm/500 nm and a fluorescence efficiency (Sf2) of a calcium dissociation dye at an excitation wave length of 380 nm/500 nm.
Calculation Formula: Intracellular Calcium Concentration
(nM)224xc3x97[(Rxe2x88x92Rmin)/(Rmaxxe2x88x92R)]xc3x97[Sf2/Sb2]
Using the thus calculated intracellular calcium concentration in the presence of a predetermined concentration of each of the drugs and that of the control solvent, a ratio of inhibiting calcium influx (CRACC inhibition) was obtained Lo calculate its concentration to inhibit 50% of CRACC (IC50 value).
(2) Selectivity of CRACC Inhibition Against VOCC
A suspension of rat neuroblasts PC12-h5 (2xc3x97106/ml) loaded with a calcium indicator fluorescence dye fura-2 (1 xcexcM) was dispensed in 100 xcexcl portions into wells of a 96 well microplate. Intracellular calcium increase stimulated with high concentration potassium chloride was induced by adding to each well a 100 xcexcl of Hanks"" balanced salt solution containing a drug to be tested in two times higher concentration than the final concentration and 100 mM of KCl (final concentration, 50 mM), and, after 30 minutes of the addition, a fluorescence intensity ratio (R) was calculated from two fluorescence intensities obtained at excitation wave lengths of 340 nm/500 nm and 380 nm/500 nm, respectively. In calculating R, self-fluorescence of the drug to be tested was measured in a cell-free system, and the effect of the self-fluorescence on the fura-2 fluorescence was corrected.
The IC50 value of VOCC inhibition was calculated in the same manner as the case of the aforementioned CRACC inhibition, and compared with that of CRACC inhibition.
The CRACC inhibition activity (IC50 value) of the novel compounds of Examples 1, 5, 32, 36, 38, 50, 53 and 72 and the known compounds A and D (both purchased from MYBRIDGE) was within the range of from 0.51 to 0.050 xcexcM. In addition, the CRACC inhibition activity of these compounds was superior to the VOCC inhibition activity by a factor of from 16 to 200, thus showing selectivity.
(3) Inhibitory Effect on IL-2 Production
Inhibitory effect of the invention compound on IL-2 production from Jurkat cells was tested in accordance with the method described by S. Clare Chung et al. in Br. J. Pharmacol., 113: 861-868, 1994, and its IC50 value was calculated.
Compounds of the Examples 1, 5, 32, 36, 38, 50, 53 and 72 and Compounds A and D showed IC50 values of 1 xcexcM or less.
(4) Effect on TNCB-induced Contact Hypersensitivity Model
In five-week-old male ICR mice (SLC), effect of the invention compound on TNCB-induced contact hypersensitivity was tested in almost the same manner as the method described in Current Protocols in Immunology (John Wiley and Sons, Inc., 1994). Compounds of this invention inhibited TNCB-induced contact hypersensitivity in a dose-dependent manner.
(5) Inhibitory Effect on Concanavalin A (ConA)-induced Hepatitis in Mice
In four to five-week-old female Balb/c mice (SLC), this test was carried out by employing a method similar to the method reported by G. Tiegs et al. in J. Clin. Invest., 90: 196-203 (1992). Compounds of this invention inhibited ConA-induced hepatitis in a dose-dependent manner.
(6) Inhibitory Effect on Collagen-induced Arthritis in Mice
In five-week-old male DBA/1J mice (Charles River Japan), inhibitory effect on arthritis was tested in he similar manner as the methods reported by Fumio Nishikaku and Yoshihiko Koga in Immunopharmacology, 25, 65-74 (1993) and by Fuminori Kato, Masanao Nomura and Kyoko Nakamura in Annals of the Rheumatic Disease, 55, 535-539 (1996). Compounds of this invention showed significant inhibition on arthritis.
(7) Inhibitory Effect on Antigen-induced Airway Eosinophilia in Rat
In four-week-old male BN rats (Charles River, Japan), inhibitory effect on antigen-induced airway eosinophilia was tested in almost the same manner as the method reported by W. Elwood et al. in Inflamm. Res., 44: 83-86 (1995). In this connection, the drug was administered 30 minutes before the antigen exposure in the case of intravenous injection or 1 hour before and 3 hours after the antigen exposure in the case of oral administration.
Tn this model, compounds of this invention inhibited numbers of infiltrated total leukocytes and that of infiltrated eosinophils into airways.
A pharmaceutical composition which contains the compound (Ixe2x80x2) of the present invention or a salt thereof and a pharmaceutically acceptable carrier can be prepared by a usually used method using at least one of compounds represented by the general formula (Ixe2x80x2) or salts thereof and a carrier or medicinal use, a filer and other additives usually used in pharmaceutical preparations. Its administration may be effected either by oral admistration in the form of tablets, pills, capsules, granules, powders, solutions and the like or by parenteral administration in the form of intravenous, intramuscular and the like injections, suppositories, percutaneous absorption preparations and the like.
The solid composition for use in the oral administration according to the present invention is used in the form of tablets, powders, granules and the like. In such a solid composition, one or more active substances are mixed with at least one inert diluent such as lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinyl pyrrolidone or aluminum magnesium silicate. By the usual procedures, the composition may contain other additives than the inert diluent, such as a lubricant (e.g., magnesium stearate or the like), a disintegrating agent (e.g., calcium cellulose glycolate or the like), a stabilizing agent (e.g., lactose or the like) and a solubilization assisting agent (e.g., glutamic acid, aspartic acid or the like). If necessary, tablets or pills may be coated with films of a sugar or a gastric or enteric substance such as sucrose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose phthalate or the like.
The liquid composition for oral administration use includes pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs and the like and contains a generally used inert diluent such as purified water or ethanol. In addition to the inert diluent, this composition may also contain auxiliary agents such as a moistening agent, a suspending agent and the like, as well as sweeteners, flavors, aromatics and antiseptics.
The injections for parenteral administration use include aseptic aqueous or non-aqueous solutions, suspensions and emulsions. Examples of the diluent for use in the aqueous solutions and suspensions include distilled water for injection use and physiological saline. Examples of the diluent for use in the non-aqueous solutions and suspensions include propylene glycol, polyethylene glycol, plant oil (e.g., olive oil or the like), alcohol (e.g., ethanol and the like), and polysorbate 80. Such a composition may further contain auxiliary agents such as an antiseptic, a moistening agent, an emulsifying agent, a dispersing agent, a stabilizing agent (lactose for example) and a solubilizarion assisting agent (glutamic acid or asoartic acid for example). These compositions are sterilized by filtration through a bacteria retaining filter, blending of a germicide or irradiation. Alternatively, they may be used by firstly making into sterile solid compositions and then dissolving them in sterile water or a sterile solvent for injection use prior to their use.
In the case of oral administration, suitable daily dose is usually from about 0.001 to 10 mg/kg body weight, and the daily dose is administered once a day or divided into 2 to 4 doses per day. In the case of intravenous injection, suitable daily dose is usually from about 0.0001 to 1 mg/kg body weight, and the daily dose is administered once per several days, or once a day or divided into a plurality of doses per day. The close is optionally decided by taking into consideration symptoms, age, sex and the like of each patient to be treated.